LC and LC-MS Separation of Peptides on Macrocyclic Glycopeptide Stationary Phases: Diastereomeric Series and Large Peptides

Soukup-Hein, Renee J.; Schneiderheinze, Jeff; Mehelic, Paul; Armstrong, Daniel W.

doi:10.1365/s10337-007-0387-3

Cited by 19 publications

(26 citation statements)

References 33 publications

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“…Single amino acid substitutions (including chiral residue substitution) were resolved. A series of enkephalin-related peptides with different numbers of D-residues have been separated using this CSP [69]. Those with three or four Damino acids were most retained by the CSP column, with YdAGFdL eluted first among six enkephalin peptides (Fig.…”

Section: Chiral Separationsmentioning

confidence: 99%

“…Enantiomeric pair E0 and E4 and diastereomeric pair E1a and E3were not separated on the non-chiral column, whereas teicoplanin-based CSP was able to resolve every peptide. Adapted with permission from Springer [69]. Electropherogram of a ventral abdominal neuron spiked with synthetic NWFa standard, which migrates faster than the endogenous NdWFa peak.…”

Section: Future Directionsmentioning

confidence: 99%

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Analysis of endogenous D-amino acid-containing peptides in Metazoa

Bai¹,

Sheeley²,

Sweedler³

2009

Frontiers of Bioanalytical Chemistry

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Peptides are chiral molecules with their structure determined by the composition and configuration of their amino acid building blocks. The naturally occurring amino acids, except glycine, possess two chiral forms. This allows the formation of multiple peptide diastereomers that have the same sequence. Although living organisms use L-amino acids to make proteins, a group of D-amino acidcontaining peptides (DAACPs) has been discovered in animals that have at least one of their residues isomerized to the D-form via an enzyme-catalyzed process. In many cases, the biological functions of these peptides are enhanced due to this structural conversion. These DAACPs are different from those known to occur in bacterial cell wall and antibiotic peptides, the latter of which are synthesized in a ribosome-independent manner. DAACPs have now also been identified in a number of distinct groups throughout the Metazoa. Their serendipitous discovery has often resulted from discrepancies observed in bioassays or in chromatographic behavior between natural peptide fractions and peptides synthesized according to a presumed all-L sequence. Because this L-to-D post-translational modification is subtle and not detectable by most sequence determination approaches, it is reasonable to suspect that many studies have overlooked this change; accordingly, DAACPs may be more prevalent than currently thought. Although diastereomer separation techniques developed with synthetic peptides in recent years have greatly aided in the discovery of natural DAACPs, there is a need for new, more robust methods for naturally complex samples. In this review, a brief history of DAACPs in animals is presented, followed by discussion of a variety of analytical methods that have been used for diastereomeric separation and detection of peptides.

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Section: Chiral Separationsmentioning

confidence: 99%

Section: Future Directionsmentioning

confidence: 99%

Analysis of endogenous D-amino acid-containing peptides in Metazoa

Bai¹,

Sheeley²,

Sweedler³

2009

Frontiers of Bioanalytical Chemistry

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“…Typically alcohol/water mobile phases were used for the separation of: enantiomers of protein amino acids [33, 39, 40, 70 -73], other unusual amino acids [70, 72, 74 -81], N-blocked amino acids [33,34,39,72,73,82]; stereoisomers (both enantiomers and diastereoisomers) of di-and tripeptides [39,70]; and structurally related peptides (diastereomers, and peptides that differ by one or a few amino acid residues) with up to 30 amino acid units [83,84].…”

Section: Separation Of Amino Acids and Peptides On Macrocyclic Glycopmentioning

confidence: 99%

“…The water content, ionic strength, and the pH of the mobile phase can be optimized by following the rationale discussed above for amino acids. Reverse gradients can be used when separating mixtures of peptides with a wide range of retention times [84]. As on many other stationary phases, the retention of amino acids and peptides on Chirobiotic CSPs produces a "U-shape" curve when retention is plotted against the organic content of the mobile phase [83].…”

Section: Mobile Phase Selectionmentioning

confidence: 99%

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Considerations on HILIC and polar organic solvent‐based separations: Use of cyclodextrin and macrocyclic glycopetide stationary phases

Wang

Jiang

Armstrong³

2008

J of Separation Science

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There is a natural tendency in science to prefer straightforward, logical classification systems. The use of mobile phase-stationary phase combinations that do not fit neatly into the standard "normal phase" or "reversed-phase" categories has been going on for over 50 years. The term "hydrophilic interaction chromatography" (HILIC) is sometimes being used as a general category for these "other" separations. In some cases, it may be appropriate and in others, not. Indeed the mechanistic constrains used to define the method seem to be varying with time. Given the name HILIC, it is assumed that water is not only present in the mobile phase, but also plays an essential role in the retention mechanism. However, there is residual water present in all organic solvents. Regardless, the number of reported separations in this alternative mode has increased tremendously in the last two decades. This is due to the advent of new stationary phases and an emphasis on polar, biologically important molecules. We discuss the relationships between HILIC and other chromatographic modes. We then examine two classes of stationary phases that have played a major role in these separations. These particular stationary phases can be used to provide appreciable mechanistic information as well.

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Separation of Peptide Diastereomers

Scriba

2014

Encyclopedia of Analytical Chemistry

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Diastereomeric peptides, isomers in which one or more of the chiral centers have been converted to the opposite configuration (R or S, D or L ), often possess different biological activities and/or conformational properties. Therefore, the separation and determination of diastereomeric peptides is important in life sciences and particularly significant to the pharmaceutical industry for quality control of peptide synthesis and stability as well as for regulatory requirements. Diastereomers differ in their physicochemical properties that can be utilized for their determination by numerous techniques. In contrast to many other methods such as optical rotation, circular dichroism, differential scanning calorimetry, or infrared (IR) spectroscopy, separation techniques such as chromatography or capillary electrophoresis (CE) allow the sensitive and simultaneous identification and quantification of peptide diastereomers. This chapter covers analytical‐scale separation of peptide diastereomers by chromatographic methods, including paper chromatography (PC), thin‐layer chromatography (TLC), gas chromatography (GC), high‐performance liquid chromatography (HPLC), and CE. HPLC is currently the predominantly used method for peptide diastereomer separations. However, owing to the high resolving power, particularly for polar compounds, CE has been applied to the separation of diastereomeric peptides in recent years. The chapter focuses on methods employing achiral stationary phases or buffers without chiral additives for the separations, although some examples of such separations, i.e. the use of chiral columns or chiral mobile phase additives, are mentioned. Moreover, only diasteromeric peptides containing natural amino acids will be discussed.

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LC and LC-MS Separation of Peptides on Macrocyclic Glycopeptide Stationary Phases: Diastereomeric Series and Large Peptides

Cited by 19 publications

References 33 publications

Analysis of endogenous D-amino acid-containing peptides in Metazoa

Analysis of endogenous D-amino acid-containing peptides in Metazoa

Considerations on HILIC and polar organic solvent‐based separations: Use of cyclodextrin and macrocyclic glycopetide stationary phases

Separation of Peptide Diastereomers

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